Cell differentiation along the erythroid lineage arises from hematopoietic stem cells within the bone marrow. The cell surface marker CD36, which is considered an early marker of erythrocyte differentiation, is acquired as the differentiation of this lineage progresses. However, the rarity and heterogeneity of the CD36 population makes it difficult to isolate in vivo. In order to obtain CD36+ erythroid progenitor cells, CD34+ hematopoietic stem cells (HSC) are isolated and cultured in serum-free expansion media supplemented with factors formulated to selectively promote the expansion and differentiation of human erythroid progenitor cells.

Human peripheral blood CD36+ erythroid progenitor cells are derived from cultured peripheral blood CD34+ cells. First, CD34+ cells are positively selected using immunomagnetic anti-CD34 microbeads from peripheral blood mononuclear cells. The isolated CD34+ cells are cultured in StemSpan™ SFEM, a serum-free expansion media for hematopoietic cells, supplemented with SCF, EPO and IL-3. Cultured cells are harvested after 10 days. Isolated cells are characterized by flow cytometry to ensure a highly pure and viable cell population.

Cells were obtained using Institutional Review Board (IRB) approved consent forms and protocols.